ACTA VET. BRNO 2011, 80: 051–056; doi:10.2754/avb201180010051 The acaricidal effect of flumethrin, oxalic acid and amitraz against Varroa destructor in honey bee (Apis mellifera carnica) colonies Maja Ivana Smodiš Škerl, Mitja Nakrst, Lucija Žvokelj, Aleš Gregorc Agricultural Institute of Slovenia, Ljubljana, Slovenia Received February 5, 2010 Accepted June 15, 2010 Abstract During 2007 and 2008, natural mite mortality was recorded in honey bee colonies. These colonies were then treated with various acaricides against Varroa destructor and acaricide efficacies were evaluated. In 2007, experimental colonies were treated with flumethrin and/or oxalic acid and in 2008 the same colonies were treated with flumethrin, oxalic acid or amitraz. The efficacy of flumethrin in 2007 averaged 73.62% compared to 70.12% for three oxalic acid treatments. In 2008, a reduction of 12.52% in mite numbers was found 4 weeks after flumethrin application, while 4 oxalic acid applications produced significantly higher (P < 0.05) mite mortality, an average of 24.13%. Four consecutive amitraz fumigations produced a 93.82% reduction on average in final mite numbers and thus ensure normal colony development and overwintering. The study is important in order to demonstrate that synthetic acaricides should be constantly re-evaluated and the use of flumethrin at low efficacies need to be superseded by appropriate organic treatments to increase the efficacy of mite control in highly-infested colonies during the period of brood rearing. Varroa control, mite mortality, efficacy, trickling, fumigation, organic treatment The ectoparasitic mite Varroa destructor is a major pest of honey bees (Apis mellifera) worldwide (Sammataro et al. 2000) that needs to be controlled because untreated colonies die within a few years due to damage to both pupae and adult bees (Elzen et al. 2000). A high rate of mite infestation in honey bee colony and a poor colony management have important influence on the beekeeping. Regular treatment of honey bee colonies with acaricides allows productivity to be maintained, but the use of synthetic acaricides with their lipophilic and persistent characteristics can result in their residues in wax and honey (Wallner 1999). The synthetic pyrethroid acaricide flumethrin (Bayvarol®; Bayer, Germany) is the only registered treatment against V. destructor for colonies with brood in Slovenia. It is relatively non-toxic to the bees and easy to use. Mites have developed resistance to several synthetic acaricides (Lodesani et al. 1995; Milani 1999; Floris et al. 2001; Spreafico et al. 2001), so beekeepers turn to alternative treatments incorporating essential oils or organic acids (Mutinelli et al. 1997; Gregorc and Poklukar 2003). Resistance of mites to acaricides has been associated with the misuse of agricultural formulations of synthetic pyrethroids (Watkins 1997). The permanent use of varroacide strips causes increasing selection pressure for resistant mites (Milani 1999). Oxalic acid is a natural constituent of honey, and EU regulations permit its use in biological beekeeping (EU Council Regulation, No. 1804/1999). Because of its high efficacy, oxalic acid is widely used in most European countries (Charriere and Imdorf 2002). Research has been conducted into the efficacy of oxalic acid applications for controlling mites in colonies with or without brood (Imdorf et al. 1996; Brødsgaard et al. 1999). Experiments demonstrated that trickling an aqueous oxalic acid solution into the hive (Mutinelli et al. 1997; Gregorc and Planinc 2001; Gregorc and Planinc 2002) during broodless periods (Radetzki 1994; Imdorf et al. 1997) was highly effective in killing the mite, achieving a mean 98% effectiveness. When capped brood is present, efficacy after 3 Address for correspondence: Phone +386 1 280-52-41 Maja Ivana Smodiš Škerl Fax +386 1 280-52-55 Agricultural Institute of Slovenia E-mail: [email protected] Hacquetova 17, 1001 Ljubljana, Slovenia http://www.vfu.cz/acta-vet/actavet.htm 52 treatments of a 5% oxalic acid solution was approximately 95% (Mutinelli et al. 1997). In our previous experiments, 50 ml of an oxalic acid solution applied to one normally developed colony had an efficacy of 39.2% when brood was present and 99.4% without brood (Gregorc and Planinc 2001). Several compounds were used against varroa mites in honeybee colonies. Amitraz, a formamidine, was successfully used to kill varroa mites when formulated as a balsa wood strip soaked in amitraz, aerosol spray or smoking paper strip (Witherell and Herbert 1988) and the organophosphate coumaphos (Perizin®) was used to document the respective efficacies of experimentally applied acaricides (Trouiller 1998). The aim of this study was to establish the effectiveness of amitraz, flumethrin and oxalic acid treatments for controlling V. destructor in honey bee colonies. We also aimed to establish an optimal strategy for summer treatment with different acaricides in colonies with brood and for winter treatments of broodless colonies. Materials and Methods Apis mellifera carnica colonies in hives containing ten combs (41 × 26 cm) in each brood chamber and honey super were located at one site at the experimental station of the Slovenian Agricultural Institute. On 2 April 2007, metal sheets (38 × 29.8 cm) were placed on the floor of each of the hives to record the colony’s natural mite mortality. Wire screens above the sheet prevent bees from coming in contact with debris. The number of mites was recorded on 14 occasions in the pre-treatment period, and then recorded once a week after 7 August when treatments began. The number of oxalic acid treatments in each colony was determined after establishing the mite mortality before and after each treatment. The colonies received 5 ml (per comb occupied by bees) of a 2.9% oxalic acid and 31.9% sucrose aqueous solution (w/w), using oxalic acid dihydrate (Riedel-de Haën), sucrose and de-mineralised water (Gregorc and Planinc 2001). The oxalic acid treatments were applied to the experimental colonies by trickling the oxalic acid solution over the combs in situ and squirting the bees in the brood chamber using a syringe. Respiration masks, protective glasses and rubber gloves were worn while applying the solution. The mite drop during the treatment period was recorded after each application. The outside temperatures during the July and August ranged from 28 to 31 °C, and temperatures during the treatments in the broodless period were above 5 °C in both years. The treatments of 51 colonies in 2007 were performed as follows: Group A (43 colonies) received flumethrin strips (4 strips per brood chamber) between 7 and 28 August during the brood period and 2 oxalic acid treatments in the broodless period on 26 November and 13 December; Group B (eight colonies) received 3 oxalic acid treatments on 7, 22 and 28 August during the brood period, and also 2 oxalic acid treatments on 26 November and 13 December during the broodless period. Oxalic acid treatments in broodless colonies were considered as treatments to establish the final mite drop (Gregorc and Planinc 2001). In the spring of 2008, metal sheets were placed on the floor of each hive on 13 May, and the number of mites was recorded on 11 occasions in the pre-treatment period and then regularly after the start of treatments on 25 July. The treatment groups in 2008 were different from the previous year and were as follows: Group A (28 colonies) received flumethrin between 25 July and 21 August; Group B (18 colonies) received four oxalic acid treatments 25 and 31 July and 7 and 14 August. After receiving flumethrin or oxalic acid, both groups of colonies were treated with amitraz. Each treated colony was fumigated using 0.1 ml (3 drops) of 12.5% emulsifiable concentrate amitraz (Tactik®) on 21 August, 11 and 22 September and 7 October. The efficacy of the flumethrin and oxalic acid treatments, expressed as % mite mortality, was established after each treatment, and the mean mite drop values of the treated groups were compared. A standard treatment with coumaphos (Perizin® 1 ml and 15 g sugar in 50 ml water; Bayer; 32 mg coumaphos/1 ml Perizin®) was conducted on 4 and 11 November 2008 in order to establish the final mite drop (Trouiller 1998). To estimate the percentage of mites killed (PMK) by the first flumethrin or oxalic acid application (PMK1) when brood was present, the following formula (Gregorc and Planinc 2001) was used: PMK1 = (T1 / (T1 + T2 + T3 + ..... + P) × 100) % T1, T2 and T3 denote the total number of mites that dropped after the first, second and third treatments respectively, including the flumethrin treatment (Group A), and P denotes the number of mites collected after the coumaphos treatment. The formula was modified to calculate the percentage of mites dropped after each consecutive treatment (PMK-T2 – PMK-P) by excluding the number of mites collected in the previous treatments. The efficacy of the treatments was also estimated by comparing the numbers of mites that fell before and after the treatments and the mite mortality between the consecutive oxalic acid treatments and between groups A and B. The data analyses were performed by ANOVA (analysis of variance) with the use of the Statgraphic (20) program. 53 Results During the pre-treatment periods, a total of 127 days at intervals between 2 April and 7 August 2007, the average daily natural mite drop was estimated to be 0.11 (± 0.14). The average mite mortality per colony during the pre-treatment period was estimated to be 13.92 (± 18.01) mites. In this period 1.12% (± 0.86 %) of the total mite population found during the experiment died naturally. Mite mortality after flumethrin application in the period from 7 to 22 August was higher (P < 0.05) compared to natural mite mortality in the pre-treatment period. The increase in mite mortality (P < 0.05) compared to pre-treatment period was found also in colonies after three oxalic acid applications.